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In vivo BrdU Incorporation and Detection in Mouse

Zheng Liu Zheng Liu

Published: Jun 5, 2011 DOI: 10.21769/BioProtoc.81 Views: 37102

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Abstract

BrdU (Bromodeoxyuridine or 5-bromo-2’-deoxyuridine) is a synthetic nucleoside that is incorporated into DNA by proliferating cells. This protocol is to be used to incorporate and detect BrdU in murine plasma cells. The plasma cells described in this protocol are formed spontaneously in autoimmune mice (NZB/W mice). Modifications are most likely needed if users intend to label plasma cells in immunized mice.

Keywords: Autoimmune search Mouse search Plasma cells search

Materials and Reagents

  1. Antibodies
    1. BD mouse Fc Block (BD Biosciences, Pharmingen™, catalog number: 553142 )
    2. Rat anti-mouse B220 APC (Southern Biotech, catalog number: 1665-11 )
    3. Rat anti-mouse CD138 PE (BD Biosciences, Pharmingen™, catalog number: 561070 )
      Note: The above antibodies have been tested by the author and may be substituted with the antibodies desired by users.

  2. Other materials
    1. Mice
    2. FITC BrdU Flow Kit (BD Biosciences, Pharmingen™, catalog number: 559619 )
      Note: *Provided in the kit.
    3. BrdU (Sigma-Aldrich, catalog number: B5002 )
    4. 1x Dulbecco’s modified eagle medium (DMEM) (Life Technologies, Invitrogen™, catalog number: 10313-039 )
    5. Fetal bovine serum (FBS) (Life Technologies, Invitrogen™, catalog number: 11091-148)
    6. Ammonium chloride 10x lysing solution (see Recipes)
    7. FACS buffer (see Recipes)

Equipment

  1. BD LSR II flow cytometer

Procedure

Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.

Category

Immunology > Immune cell function > General

Developmental Biology > Cell growth and fate

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